CN110372105B - Compound microbial agent for improving aquaculture water and preparation method thereof - Google Patents
Compound microbial agent for improving aquaculture water and preparation method thereof Download PDFInfo
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- CN110372105B CN110372105B CN201910624847.XA CN201910624847A CN110372105B CN 110372105 B CN110372105 B CN 110372105B CN 201910624847 A CN201910624847 A CN 201910624847A CN 110372105 B CN110372105 B CN 110372105B
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 66
- 230000000813 microbial effect Effects 0.000 title claims abstract description 43
- 239000003795 chemical substances by application Substances 0.000 title claims abstract description 39
- 238000009360 aquaculture Methods 0.000 title claims abstract description 38
- 244000144974 aquaculture Species 0.000 title claims abstract description 38
- 150000001875 compounds Chemical class 0.000 title claims abstract description 27
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 238000000855 fermentation Methods 0.000 claims abstract description 50
- 230000004151 fermentation Effects 0.000 claims abstract description 50
- 241000894006 Bacteria Species 0.000 claims abstract description 15
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 14
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 11
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 11
- 241000193752 Bacillus circulans Species 0.000 claims abstract description 9
- 241000194107 Bacillus megaterium Species 0.000 claims abstract description 9
- 241000235646 Cyberlindnera jadinii Species 0.000 claims abstract description 9
- 241000194032 Enterococcus faecalis Species 0.000 claims abstract description 9
- 240000001046 Lactobacillus acidophilus Species 0.000 claims abstract description 9
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 claims abstract description 9
- 241000190950 Rhodopseudomonas palustris Species 0.000 claims abstract description 9
- 229940039695 lactobacillus acidophilus Drugs 0.000 claims abstract description 9
- 241000108664 Nitrobacteria Species 0.000 claims abstract description 8
- 239000000047 product Substances 0.000 claims abstract description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N glycerol group Chemical group OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 33
- 239000007788 liquid Substances 0.000 claims description 25
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 18
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 18
- 238000004108 freeze drying Methods 0.000 claims description 17
- 239000003223 protective agent Substances 0.000 claims description 17
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 14
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- 241000237502 Ostreidae Species 0.000 claims description 12
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 235000020636 oyster Nutrition 0.000 claims description 12
- 239000000843 powder Substances 0.000 claims description 12
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 10
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 10
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 241000238557 Decapoda Species 0.000 claims description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- 229930006000 Sucrose Natural products 0.000 claims description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 6
- 239000012153 distilled water Substances 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 238000005286 illumination Methods 0.000 claims description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 6
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 6
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 6
- 239000002244 precipitate Substances 0.000 claims description 6
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 230000001580 bacterial effect Effects 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 3
- 238000011081 inoculation Methods 0.000 claims description 3
- 230000000844 anti-bacterial effect Effects 0.000 claims description 2
- 239000003899 bactericide agent Substances 0.000 claims description 2
- 230000001737 promoting effect Effects 0.000 claims description 2
- 239000012629 purifying agent Substances 0.000 claims description 2
- 241001465754 Metazoa Species 0.000 abstract description 16
- 239000000126 substance Substances 0.000 abstract description 11
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 7
- 239000001301 oxygen Substances 0.000 abstract description 7
- 229910052760 oxygen Inorganic materials 0.000 abstract description 7
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 abstract description 5
- 244000005700 microbiome Species 0.000 abstract description 5
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 abstract description 4
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 abstract description 4
- 229910000037 hydrogen sulfide Inorganic materials 0.000 abstract description 4
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 abstract description 3
- 244000052616 bacterial pathogen Species 0.000 abstract description 3
- 230000008901 benefit Effects 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 229910052698 phosphorus Inorganic materials 0.000 abstract description 3
- 239000011574 phosphorus Substances 0.000 abstract description 3
- 230000000295 complement effect Effects 0.000 abstract description 2
- 239000013586 microbial product Substances 0.000 abstract description 2
- 235000015097 nutrients Nutrition 0.000 abstract description 2
- 230000000644 propagated effect Effects 0.000 abstract description 2
- 230000000694 effects Effects 0.000 description 7
- 239000003607 modifier Substances 0.000 description 7
- 229960004793 sucrose Drugs 0.000 description 5
- 240000008042 Zea mays Species 0.000 description 4
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 4
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 235000005822 corn Nutrition 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000194108 Bacillus licheniformis Species 0.000 description 2
- 230000005784 autoimmunity Effects 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 239000003640 drug residue Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000009777 vacuum freeze-drying Methods 0.000 description 2
- 238000003911 water pollution Methods 0.000 description 2
- 241000222122 Candida albicans Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- GQPLMRYTRLFLPF-UHFFFAOYSA-N Nitrous Oxide Chemical compound [O-][N+]#N GQPLMRYTRLFLPF-UHFFFAOYSA-N 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229940095731 candida albicans Drugs 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 230000001546 nitrifying effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000003895 organic fertilizer Substances 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
- A01K61/13—Prevention or treatment of fish diseases
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K63/00—Receptacles for live fish, e.g. aquaria; Terraria
- A01K63/04—Arrangements for treating water specially adapted to receptacles for live fish
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/101—Sulfur compounds
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/105—Phosphorus compounds
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
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- Zoology (AREA)
- Environmental Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Marine Sciences & Fisheries (AREA)
- Animal Husbandry (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Hydrology & Water Resources (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Botany (AREA)
- Farming Of Fish And Shellfish (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the field of microbial products, and particularly relates to a compound microbial agent for improving an aquaculture water body and a preparation method thereof. The microbial inoculum is a fermentation product of a mixed strain of bacillus subtilis, rhodopseudomonas palustris, bacillus megaterium, lactobacillus acidophilus, streptococcus faecalis, bacillus circulans, candida utilis, nitrobacteria and denitrifying bacteria. The compound microbial agent disclosed by the invention contains microorganisms capable of rapidly providing dissolved oxygen, microorganisms capable of decomposing ammonia nitrogen, hydrogen sulfide and nitrite in a water body, and strains capable of providing nutrients required for production and development of aquatic animals. The advantages of the strains are complementary, and the strains play a role in synergy; after being further applied to a water body, the aquatic product water body can be rapidly propagated to form a dominant population, macromolecular organic matters, residual bait, excrement and the like in the aquaculture water body can be rapidly decomposed, ammonia nitrogen, phosphorus, nitrite, hydrogen sulfide and other substances in the water body can be degraded, the water quality can be purified, the dissolved oxygen content of the water body can be improved, and pathogenic bacteria can be removed.
Description
Technical Field
The invention belongs to the field of microbial products, and particularly relates to a compound microbial agent for improving an aquaculture water body and a preparation method thereof.
Background
China is a world large aquaculture country, and in recent years, with the rapid development of aquaculture industry in China, the continuous expansion of aquaculture scale, the water pollution is more and more serious. The water body is an environment which is necessary for the survival and growth of aquatic animals such as fishes, shrimps and the like, and the quality of the water quality is the key to the success or failure of aquaculture, particularly intensive aquaculture. The aquaculture mode of high density intensification is mostly adopted in China, the culture water body pollution and the imbalance of water quality factors are easily caused by the human activities and the factors of self secretion, excrement, excessive bait, drug residue and the like of culture objects, and the aquatic product culture animals are attacked or even die in large quantities.
The water quality modifier used at present mainly comprises a physical water quality modifier, a chemical water quality modifier and a biological water quality modifier. The physical water quality modifier mainly adsorbs harmful substances rapidly through adsorption, and has slow and unobvious action effect and is not easy to degrade. The chemical water quality improver can react with harmful substances to degrade the harmful substances, but is easy to cause secondary pollution, and has irritation and adverse effects on aquatic animals, particularly seedling stage animals. The biological water quality modifier can inhibit the growth and reproduction of pathogenic bacteria, thoroughly decompose harmful substances, provide trace elements for aquatic animals, improve the autoimmunity of the aquatic animals and restore the ecological environment of water bodies.
The biological water quality modifier is a compound microbial agent, is prepared from two or more beneficial microbial bacteria which are not antagonistic to each other, and is widely applied to various fields such as aquaculture, sewage treatment and the like. However, the use of the existing microbial preparation is greatly influenced by external conditions such as environmental temperature, chemical drug residues in water and the like, the application requirement is high, and the actual effect of the microbial preparation is greatly fluctuated. The microbial strains in the microbial inoculum consume oxygen during mass growth and propagation, so that the dissolved oxygen in the water body is reduced, and metabolites in the growth process can change the water body environment and cause certain influence on aquatic animals.
Disclosure of Invention
The invention aims to provide a compound microbial agent for improving aquaculture water.
The second purpose of the invention is to provide a preparation method of the compound microbial agent.
In order to achieve the purpose, the invention provides the following technical scheme:
a composite microbial agent for improving aquaculture water is prepared from Bacillus subtilis, rhodopseudomonas palustris, bacillus megaterium, lactobacillus acidophilus, streptococcus faecalis, bacillus circulans, candida utilis, nitrifying bacteria and denitrifying bacteria.
The microbial inoculum is prepared by adding oyster powder, biological peptide and a freeze-drying protective agent into mixed strain fermentation liquor, uniformly mixing, centrifuging and collecting precipitate to obtain the compound microbial inoculum; wherein the weight ratio of the oyster powder, the biological peptide, the freeze-drying protective agent to the fermentation liquor is (1-10): (0.1-5), (10-20): 100.
the biological peptide is corn peptide; the freeze-drying protective agent is a glycerol solution and a trehalose solution in a mass ratio of 1-3 to 1-2, wherein the concentration of the glycerol solution is 30% and the concentration of the trehalose solution is 10%.
The fermentation liquor is prepared by adding the mixed strain into the base liquor of the fermentation liquor according to the inoculum size of 3-5%, and culturing for 5-8 days at 30-37 deg.C under illumination until the total bacterial density in the fermentation liquor reaches (1-10) x 10 8 Stopping fermentation at the seed/L; wherein, the fermentation liquor base liquid comprises, by weight, 30-50 parts of acetic acid, 50-70 parts of ethanol, 150-200 parts of sucrose water, 10-30 parts of magnesium sulfate, 20-50 parts of monopotassium phosphate, 50-100 parts of sodium chloride, 20-50 parts of sodium carbonate, 150-300 parts of glucose and 300-500 parts of distilled water.
The mixed strain comprises, by weight, 8-12 parts of bacillus subtilis, 10-13 parts of rhodopseudomonas palustris, 8-12 parts of bacillus megaterium, 4-7 parts of lactobacillus acidophilus, 5-8 parts of streptococcus faecalis, 8-11 parts of bacillus circulans, 7-10 parts of candida utilis, 4-9 parts of nitrobacteria and 5-8 parts of denitrifying bacteria.
Further, the base solution of the fermentation liquid comprises, by weight, 40 parts of acetic acid, 60 parts of ethanol, 180 parts of sucrose solution, 20 parts of magnesium sulfate, 35 parts of monopotassium phosphate, 75 parts of sodium chloride, 35 parts of sodium carbonate, 240 parts of glucose and 400 parts of distilled water.
A method for preparing a compound microbial agent for improving aquaculture water is obtained by adding mixed strains into a fermentation broth base solution for fermentation.
The microbial inoculum is prepared by adding oyster powder, biological peptide and a freeze-drying protective agent into mixed strain fermentation liquor, evenly mixing, centrifuging and collecting precipitate to obtain the compound microbial inoculum; wherein the weight ratio of the oyster powder, the biological peptide, the freeze-drying protective agent to the fermentation liquor is (1-10): (0.1-5), (10-20): 100.
further, the following steps are carried out:
(1) Weighing each base material according to the weight of each component in the fermentation liquid base solution, fully dissolving, adjusting the pH to 7.0, and carrying out high-pressure sterilization to prepare a fermentation liquid base solution for later use;
(2) According to the weight parts, 8-12 parts of bacillus subtilis, 10-13 parts of rhodopseudomonas palustris, 8-12 parts of bacillus megaterium, 4-7 parts of lactobacillus acidophilus, 5-8 parts of streptococcus faecalis, 8-11 parts of bacillus circulans, 7-10 parts of candida utilis, 4-9 parts of nitrobacteria and 5-8 parts of denitrifying bacteria are taken, and all strains are respectively inoculated into a fermentation base liquid to be cultured for 3-7 days under illumination at 30-37 ℃;
(3) Respectively transferring the bacterial liquids obtained in the step (2) to a fermentation base liquid again for amplification culture, and culturing for 5-8 days at 30-37 ℃ under illumination until the total bacterial density in the fermentation liquid reaches (1-10) x 10 8 Stopping fermentation at the seed/L;
(4) Adding oyster powder, biological peptide and a freeze-drying protective agent into the fermentation liquor, fully stirring and mixing for 8-12h, centrifuging by a centrifugal machine, removing supernatant, and carrying out vacuum freeze drying on the precipitate to obtain the compound microbial agent.
The freeze-drying protective agent comprises a glycerol solution and a trehalose solution in a mass ratio of 1-3 to 1-2, wherein the concentration of the glycerol solution is 30% and the concentration of the trehalose solution is 10%; wherein, the glycerol solution and the trehalose solution are all commercial products.
A preparation method of a compound microbial agent for improving aquaculture water, wherein the microbial agent is applied as a bactericide of the aquaculture water, a purifying agent of the aquaculture water or a growth promoting agent for aquaculture.
The beneficial effects of the invention are:
the compound microbial agent disclosed by the invention contains microorganisms capable of rapidly providing dissolved oxygen, microorganisms capable of decomposing ammonia nitrogen, hydrogen sulfide and nitrite in a water body, and strains capable of providing nutrients required for production and development of aquatic animals. The advantages of the strains are complementary, and the strains play a role in synergy; after being further applied to a water body, the biological organic fertilizer can be rapidly propagated to form a dominant population, so that macromolecular organic matters, residual bait excrement and the like in the aquaculture water body can be rapidly decomposed, ammonia nitrogen, phosphorus, nitrite, hydrogen sulfide and other substances in the water body can be degraded, the water quality can be purified, the dissolved oxygen content of the water body can be improved, and pathogenic bacteria, such as pathogenic escherichia coli, mould, staphylococcus aureus, candida albicans and the like can be removed. The composite microbial inoculum has good stability and high efficiency, can strengthen the constitution of aquatic animals, improve the quality of the animals and reduce the death rate in the breeding process. And the preparation method of the microbial agent is simple and the production cost is low.
The specific implementation mode is as follows:
the technical solution of the present invention is further described with reference to the following embodiments, and the described embodiments are only a part of the embodiments of the present invention, but the scope of the present invention is not limited thereto.
Example 1: a preparation method of a compound microbial agent for improving aquaculture water comprises the following steps:
(1) According to the weight of each component in the fermentation liquid base liquid, 30g of fermentation liquid base material acetic acid, 50 g of ethanol, 150 g of sucrose water, 10 g of magnesium sulfate, 20 g of monopotassium phosphate, 50 g of sodium chloride, 20 g of sodium carbonate, 150 g of glucose and 300 g of distilled water are respectively weighed. Fully dissolving, adjusting pH to 7.0, and autoclaving to obtain fermentation broth base solution;
(2) According to parts by weight, 8 g of bacillus subtilis, 10 g of rhodopseudomonas palustris, 8 g of bacillus megaterium, 4 g of lactobacillus acidophilus, 5 g of streptococcus faecalis, 8 g of bacillus circulans, 7 g of candida utilis, 4 g of nitrobacteria and 5 g of denitrifying bacteria are taken, all strains are inoculated into a fermentation base liquid according to the inoculation amount of 3 percent of the mass ratio and are cultured for 3 to 7 days under the illumination of 30 to 32 ℃, all bacteria liquid is inoculated into the fermentation base liquid again according to the inoculation amount of 3 percent of the mass ratio and are transferred to the fermentation base liquid for expanding culture, the bacteria liquid is cultured for 5 to 8 days under the illumination of 30 to 32 ℃ until the total bacteria density in the fermentation liquid reaches (8 to 10) multiplied by 10, and the total bacteria density in the fermentation liquid reaches (8 to 10) 8 Stopping fermentation at the time of seed/L.
(3) The weight ratio of the oyster powder, the corn peptide, the freeze-drying protective agent to the fermentation liquor is 3:1:10:100, wherein the freeze-drying protective agent is a 30% glycerol solution (namely 30g of glycerol dissolved in 70g of water) and a 10% trehalose solution in mass percentage, and the mass ratio of the two is 3. Fully stirring and mixing for 10h, centrifuging at 4000rpm/min, removing supernatant, pre-freezing the precipitate for 5h at-50 ℃, gradually heating to 30 ℃, drying for 12h totally, and performing vacuum freeze drying to obtain the compound microbial agent.
Example 2:
the complex microbial preparation was prepared according to the procedure described in example 1, except that:
(1) According to the weight of each component in the fermentation liquid base liquid, 40 g of fermentation liquid base material acetic acid, 60 g of ethanol, 180 g of cane sugar water, 20 g of magnesium sulfate, 35 g of monopotassium phosphate, 75 g of sodium chloride, 35 g of sodium carbonate, 240 g of glucose and 400 g of distilled water are respectively weighed. Fully dissolving, adjusting pH to 7.0, and autoclaving to obtain fermentation broth base solution;
(2) According to the weight portion, 10 g of bacillus subtilis, 12 g of rhodopseudomonas palustris, 10 g of bacillus megaterium, 5 g of lactobacillus acidophilus, 7 g of streptococcus faecalis, 9 g of bacillus circulans, 8 g of candida utilis, 7 g of nitrobacteria and 6 g of denitrifying bacteria are taken.
(3) The weight ratio of the oyster powder, the corn peptide, the freeze-drying protective agent to the fermentation liquor is 5:3:15:100 (embodiment with specific point value)
Example 3:
the complex microbial preparation was prepared according to the procedure described in example 1, except that:
(1) According to the weight of each component in the fermentation liquid base liquid, 50 g of fermentation liquid base material acetic acid, 70g of ethanol, 200 g of cane sugar water, 30g of magnesium sulfate, 50 g of monopotassium phosphate, 100 g of sodium chloride, 50 g of sodium carbonate, 300 g of glucose and 500 g of distilled water are respectively weighed. Fully dissolving, adjusting pH to 7.0, and autoclaving to obtain fermentation broth base solution;
(2) According to the weight portion, 12 g of bacillus subtilis, 13 g of rhodopseudomonas palustris, 12 g of bacillus megaterium, 7 g of lactobacillus acidophilus, 8 g of streptococcus faecalis, 11 g of bacillus circulans, 10 g of candida utilis, 9 g of nitrobacteria and 8 g of denitrifying bacteria are taken.
(3) The weight ratio of the oyster powder, the corn peptide, the freeze-drying protective agent to the fermentation liquor is 9:5:20:100.
example 4: two 20-mu prawn culture ponds are selected by using the compound microbial agent special for aquatic animals prepared by the method, and the microbial agent in the embodiment 2 is put into one pond for application test, wherein the adding amount is 1kg; the other was not dosed as a control and the results are shown in table 1:
table 1: application test of aquatic animals
Compared with a control group, the test group has obvious degradation effects on Chemical Oxygen Demand (COD), ammonia nitrogen (NH 4+ -N) and nitroso nitrogen (NO 2-N) in the pond, the release period of the product is 1-2 months after the compound microbial agent is put in, and after 1 month, all indexes of the water body of the aquaculture pond are stable and the water quality is good. Beneficial microorganisms in the compound microbial agent are metabolized to generate substances which are beneficial to the growth of aquatic animals in the growth process, the effect of the compound microbial agent not only greatly improves the water quality, further enhances the autoimmunity and intestinal microenvironment of the aquatic animals, but also inhibits the propagation of pathogenic microorganisms in water, wherein the disease incidence of test groups of prawns is reduced by 54% compared with that of control groups, the yield of the prawns is relatively improved by 30%, and the economic benefit of aquaculture is improved.
Meanwhile, the compound microbial agent obtained in the embodiment is applied to the aquaculture water body, and the effect is also achieved.
Comparative example 1: microbial agent for aquaculture produced by certain biotechnology limited company
The method comprises the following steps: 2L of 5 parts of aquaculture water is taken, 2g of the microbial inoculum (mainly containing bacillus subtilis and bacillus licheniformis) for aquaculture produced in the embodiment 2 and a certain biological technology company is added respectively, the microbial inoculum is added once every three days, and the result is measured after three weeks of continuous feeding.
The content of each component in the water body before the test is shown in Table 1
TABLE 1
| COD(mg/L) | NH 4 + -N(mg/L) | Total nitrogen (mg/L) | Total phosphorus (mg/L) | NO 2 - -N(mg/L) |
| 193 | 9.5 | 17.4 | 2.07 | 1.88 |
The specific conditions of the water body after adding the microbial inoculum are shown in Table 2
TABLE 2
The comparison shows that the microbial agent produced by the invention effectively solves the problem of water pollution in aquaculture and is not easy to generate secondary pollution.
Comparative example 2: microbial agent for aquaculture produced by certain biotechnology limited company
The method comprises the following steps: selecting 4 20 mu prawn culture ponds, respectively applying 1000g of microbial agents prepared in the embodiment 2 and the comparative example (microbial agents for aquaculture, which are produced by a certain biotechnology limited company and contain bacillus subtilis and bacillus licheniformis), wherein one microbial agent is not added, the microbial agent is applied once every 15 days, and the result is measured after three months of continuous feeding.
| Detecting items | Example 2 | Comparative example | Group without addition of microbial inoculum |
| Average weight of prawn g | 30.4 | 24.7 | 20.3 |
The comparison shows that the microbial agent produced by the invention improves the growth condition of aquatic animals and improves the yield and quality of aquatic products. Is a safe and efficient microbial agent for aquaculture, and is worth of popularization and use.
The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention, and all the changes or substitutions should be covered within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
Claims (4)
1. A compound microbial agent for improving aquaculture water is characterized in that: the microbial inoculum is a compound microbial inoculum prepared by adding oyster powder, biological peptide and a freeze-drying protective agent into a mixed strain fermentation liquor, uniformly mixing, centrifuging and collecting precipitate; wherein the mass ratio of the oyster powder, the biological peptide, the freeze-drying protective agent and the fermentation liquor is (1) - (10): 0.1) - (5): 10) - (20): 100; the mixed strain is a mixed strain of bacillus subtilis, rhodopseudomonas palustris, bacillus megaterium, lactobacillus acidophilus, streptococcus faecalis, bacillus circulans, candida utilis, nitrobacteria and denitrifying bacteria;
the aquatic product is prawn;
the freeze-drying protective agent is a glycerol solution and a trehalose solution in a mass ratio of 1-3; wherein, the concentration of the glycerol solution is 30 percent, and the concentration of the trehalose solution is 10 percent;
the fermentation liquor is prepared by adding the mixed strain into a fermentation liquor base liquor according to the inoculation amount of 3-5%, and culturing for 5-8 days at 30-37 ℃ under illumination until the total bacterial density in the fermentation liquor reaches (1 to 10) multiplied by 10 8 Stopping fermentation at the seed/L; wherein, the fermentation liquor base liquid comprises, by weight, 30-50 parts of acetic acid, 50-70 parts of ethanol, 150-200 parts of sucrose water, 10-30 parts of magnesium sulfate, 20-50 parts of monopotassium phosphate, 50-100 parts of sodium chloride, 20-50 parts of sodium carbonate, 150-300 parts of glucose and 300-500 parts of distilled water;
the mixed strain comprises, by weight, 8-12 parts of bacillus subtilis, 10-13 parts of rhodopseudomonas palustris, 8-12 parts of bacillus megaterium, 4-7 parts of lactobacillus acidophilus, 5-8 parts of streptococcus faecalis, 8-11 parts of bacillus circulans, 7-10 parts of candida utilis, 4-9 parts of nitrobacteria and 5-8 parts of denitrifying bacteria.
2. The preparation method of the compound microbial agent for improving the aquaculture water body as claimed in claim 1, wherein the preparation method comprises the following steps: adding the mixed strain into a fermentation broth base solution for fermentation to obtain a fermentation broth, adding oyster powder, biological peptide and a freeze-drying protective agent into the fermentation broth, uniformly mixing, and centrifuging and collecting precipitate to obtain the compound microbial agent; wherein the mass ratio of the oyster powder, the biological peptide, the freeze-drying protective agent and the fermentation liquor is (1) - (10): 0.1) - (5): 10) - (20): 100.
3. The method for preparing the compound microbial agent for improving the aquaculture water body according to claim 2, which is characterized in that: the freeze-drying protective agent is a glycerol solution and a trehalose solution in a mass ratio of 1-3; wherein, the concentration of the glycerol solution is 30 percent, and the concentration of the trehalose solution is 10 percent.
4. The use of the compound microbial agent for improving aquaculture water according to claim 1, wherein: the microbial inoculum is applied as a bactericide of an aquaculture water body, a purifying agent of the aquaculture water body or a growth promoting agent of aquaculture.
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| CN111925964B (en) * | 2020-08-21 | 2022-04-22 | 扬州大学 | Bacillus megaterium and application thereof |
| CN113355316A (en) * | 2021-06-08 | 2021-09-07 | 聚芯生物工程有限公司 | Embedded compound microbial preparation and preparation method thereof |
| CN114045228B (en) * | 2021-09-27 | 2024-03-26 | 广州先进技术研究所 | Microbial water quality regulator and preparation method and application thereof |
| CN114292762B (en) * | 2021-12-31 | 2023-06-23 | 青岛蔚蓝赛德生物科技有限公司 | Candida palmi and application thereof |
| CN118126881A (en) * | 2023-05-26 | 2024-06-04 | 江苏海洋大学 | Heterotrophic nitrifying bacteria JHD-Z1 and microbial inoculum, preparation method and application thereof |
| CN116924848A (en) * | 2023-07-28 | 2023-10-24 | 康锌生物科技(北京)有限公司 | Composite bacterial fertilizer, preparation method thereof and application thereof in aquaculture |
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